This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gerard, G F Articles by Grandgenett, D P Search for Related Content PubMed PubMed Citation Articles by Gerard, G F Articles by Grandgenett, D P

 Previous Article  |  Next Article 

J Virol. 1975 April; 15(4): 785-797

Purification and characterization of the DNA polymerase and RNase H activities in Moloney murine sarcoma-leukemia virus.

ABSTRACT

Two RNase H (RNA-DNA hybrid ribonucleotidohydrolase, EC 3.1.4.34) activities separable by Sephadex G-100 gel filtration were identified in lysates of Moloney murine sarcoma-leukemia virus (MSV). The larger enzyme, which we have called RNase H-I, represented about 10% of the RNase H activity in the virion. RNase H-I (i) copurified with RNA-directed DNA polymerase from the virus, (ii) had a sedimentation coefficient of 4.4S (corresponds to an apparent mol wt of 70,000), (iii) required Mn-2+ (2 mM optimum) for activity with a [3-h]poly(A)-poly(dT) substrate, (iv) eluted from phosphocellulose at 0.2 M KC1, and (v) degraded [3-H]poly(A)-poly(dT) and [3-H]poly(C)-poly(dG) at approximately equal rates. The smaller enzyme, designated RNase H-II, which represented the majority of the RNase H activity in the virus preparation, was shown to be different since it (i) had no detectable, associated DNA polymerase activity, (ii) had a sedmimentation coefficient of 2.6S (corresponds to an apparent mol wt of 30,000), (iii) preferred Mg-2+ (10 to 15 mM optimum) over Mn-2+ (5 to 10 mM optimum) 2.5-fold for the degradation of [3-H]poly(A)-poly(dT), and (iv) degraded [3-H]poly(A)-poly(dT) 6 and 60 times faster than [3-H]poly(C)-poly(dG) in the presence of Mn-2+ and Mg-2+, respectively. Moloney MSV DNA polymerase (RNase H-I), purified by Sephadex G-100 gel filtration followed by phosphocellulose, poly(A)-oligo(dT)-cellulose, and DEAE-cellulose chromatography, transcribed heteropolymeric regions of avian myeloblastosis virus 70S RNA at a rate comparable to avian myeloblastosis virus DNA polymerase purified by the same procedure.

This article has been cited by other articles:

• Basavapathruni, A., Anderson, K. S. (2007). Reverse transcription of the HIV-1 pandemic. FASEB J. 21: 3795-3808 [Abstract] [Full Text]  
• Gerard, G. F., Potter, R. J., Smith, M. D., Rosenthal, K., Dhariwal, G., Lee, J., Chatterjee, Deb. K. (2002). The role of template-primer in protection of reverse transcriptase from thermal inactivation. Nucleic Acids Res 30: 3118-3129 [Abstract] [Full Text]  
• Vartanian, J.-P., Sala, M., Henry, M., Wain-Hobson, S., Meyerhans, A. (1999). Manganese cations increase the mutation rate of human immunodeficiency virus type 1 ex vivo. J. Gen. Virol. 80: 1983-1986 [Abstract] [Full Text]  
• Trentin, B., Rebeyrotte, N., Mamoun, R. Z. (1998). Human T-Cell Leukemia Virus Type 1 Reverse Transcriptase (RT) Originates from the pro and pol Open Reading Frames and Requires the Presence of RT-RNase H (RH) and RT-RH-Integrase Proteins for Its Activity. J. Virol. 72: 6504-6510 [Abstract] [Full Text]